Leishmania spp. parasite isolation through inoculation of patient biopsy macerates in interferon gamma knockout mice

Isolation of Leishmania parasite and species identification are important for confirmation and to help define the epidemiology of the leishmaniasis. Mice are often used to isolate pathogens, but the most common mouse strains are resistant to infection with parasites from the Leishmania (Viannia) subgenus. In this study we tested the inoculation of interferon gamma knockout (IFNγ KO) mice with biopsy macerates from Leishmania-infected patients to increase the possibility of isolating parasites. Biopsies from twenty five patients with clinical signs of leishmaniasis were taken and tested for the presence of parasites. Immunohistochemical assay (IHC) and conventional histopathology detected the parasite in 88% and 83% of the patients, respectively. Leishmania sp. were isolated in biopsy macerates from 52% of the patients by culture in Grace's insect medium, but 13% of isolates were lost due to contamination. Inoculation of macerates in IFNγ KO mice provides isolation of parasites in 31.8% of the biopsies. Most isolates belong to L. (Viannia) subgenus, as confirmed by PCR, except one that belongs to L. (Leishmania) subgenus. Our preliminary results support the use of IFNγ KO mice to improve the possibility to isolate New World Leishmania species.

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Naphthoquinones and derivatives for chemotherapy: perspectives and limitations of their anti-trypanosomatids activities

Current Pharmaceutical Design ◽

10.2174/1381612826666201109111802 ◽

2020 ◽

Vol 26 ◽

Author(s):

Luíza Dantas-Pereira ◽

Edézio F. Cunha-Junior ◽

Valter V. Andrade-Neto ◽

John F. Bower ◽

Guilherme A. M. Jardim ◽

...

Keyword(s):

Large Scale ◽

Neglected Tropical Diseases ◽

Folk Medicine ◽

Leishmania Species ◽

Parasitic Infections ◽

Mechanistic Analysis ◽

Leishmania Spp ◽

Nanomolar Range

: Chagas disease, Sleeping sickness and Leishmaniasis, caused by trypanosomatids Trypanosoma cruzi, Trypanosoma brucei and Leishmania spp., respectively, are considered neglected tropical diseases, and they especially affect impoverished populations in the developing world. The available chemotherapies are very limited and a search for alternatives is still necessary. In folk medicine, natural naphthoquinones have been employed for the treatment of a great variety of illnesses, including parasitic infections. This review is focused on the anti-trypanosomatid activity and mechanistic analysis of naphthoquinones and derivatives. Among all the series of derivatives tested in vitro, naphthoquinone-derived 1,2,3-triazoles were very active on T. cruzi infective forms in blood bank conditions, as well as in amastigotes of Leishmania spp. naphthoquinones containing a CF3 on a phenyl amine ring inhibited T. brucei proliferation in the nanomolar range, and naphthopterocarpanquinones stood out for their activity on a range of Leishmania species. Some of these compounds showed a promising selectivity index (SI) (30 to 1900), supporting further analysis in animal models. Indeed, high toxicity to the host and inactivation by blood components are crucial obstacles to be overcome to use naphthoquinones and/or their derivatives for chemotherapy. Multidisciplinary initiatives embracing medicinal chemistry, bioinformatics, biochemistry, and molecular and cellular biology need to be encouraged to allow the optimization of these compounds. Large scale automated tests are pivotal for the efficiency of the screening step, and subsequent evaluation of both the mechanism of action in vitro and pharmacokinetics in vivo are essential for the development of a novel, specific and safe derivative, minimizing adverse effects.

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In vitro interaction between paromomycin sulphate and four drugs with leishmanicidal activity against three New World Leishmania species

Journal of Antimicrobial Chemotherapy ◽

10.1093/jac/dkt318 ◽

2013 ◽

Vol 69 (1) ◽

pp. 150-154 ◽

Cited By ~ 25

Author(s):

E. de Morais-Teixeira ◽

M. K. Gallupo ◽

L. F. Rodrigues ◽

A. J. Romanha ◽

A. Rabello

Keyword(s):

New World ◽

Leishmania Species ◽

Leishmanicidal Activity ◽

In Vitro Interaction

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Polymerase chain reaction-based identification of New World Leishmania species complexes by specific kDNA probes

Acta Tropica ◽

10.1016/s0001-706x(99)00025-x ◽

1999 ◽

Vol 73 (3) ◽

pp. 283-293 ◽

Cited By ~ 14

Author(s):

S.F Brenière ◽

J Telleria ◽

M.F Bosseno ◽

R Buitrago ◽

B Bastrenta ◽

...

Keyword(s):

Polymerase Chain Reaction ◽

New World ◽

Leishmania Species ◽

Chain Reaction ◽

Species Complexes ◽

Polymerase Chain

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Descemet's membrane detachments, ruptures, and separations in ten adult horses: Clinical signs, diagnostics, treatment options, and preliminary results

Veterinary Ophthalmology ◽

10.1111/vop.12793 ◽

2020 ◽

Vol 23 (4) ◽

pp. 611-623

Author(s):

Roxanne M. Rodriguez Galarza ◽

Richard J. McMullen

Keyword(s):

Treatment Options ◽

Clinical Signs ◽

Preliminary Results ◽

Descemet's Membrane ◽

Descemet’S Membrane

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The NADPH Oxidase Isoform 1 Contributes to Angiotensin II-Mediated DNA Damage in the Kidney

Antioxidants ◽

10.3390/antiox9070586 ◽

2020 ◽

Vol 9 (7) ◽

pp. 586

Author(s):

Anna Zimnol ◽

Nora Spicker ◽

Ronja Balhorn ◽

Katrin Schröder ◽

Nicole Schupp

Keyword(s):

Oxidative Stress ◽

Blood Pressure ◽

Dna Damage ◽

Angiotensin Ii ◽

Knockout Mice ◽

Dna Lesions ◽

Mouse Strains ◽

Nadph Oxidases ◽

Systemic Oxidative Stress ◽

The Impact

In higher concentrations, the blood pressure regulating hormone angiotensin II leads to vasoconstriction, hypertension, and oxidative stress by activating NADPH oxidases which are a major enzymatic source of reactive oxygen species (ROS). With the help of knockout animals, the impact of the three predominant NADPH oxidases present in the kidney, i.e., Nox1, Nox2 and Nox4 on angiotensin II-induced oxidative damage was studied. Male wildtype (WT) C57BL/6 mice, Nox1-, Nox2- and Nox4-deficient mice were equipped with osmotic minipumps, delivering either vehicle (PBS) or angiotensin II, for 28 days. Angiotensin II increased blood pressure and urinary albumin levels significantly in all treated mouse strains. In Nox1 knockout mice these increases were significantly lower than in WT, or Nox2 knockout mice. In WT mice, angiotensin II also raised systemic oxidative stress, ROS formation and DNA lesions in the kidney. A local significantly increased ROS production was also found in Nox2 and Nox4 knockout mice but not in Nox1 knockout mice who further had significantly lower systemic oxidative stress and DNA damage than WT animals. Nox2 and Nox4 knockout mice had increased basal DNA damage, concealing possible angiotensin II-induced increases. In conclusion, in the kidney, Nox1 seemed to play a role in angiotensin II-induced DNA damage.

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Dynamic pelvic floor MR imaging at 3 T in patients with clinical signs of urinary incontinence-preliminary results

European Radiology ◽

10.1007/s00330-008-1030-x ◽

2008 ◽

Vol 18 (11) ◽

pp. 2620-2627 ◽

Cited By ~ 5

Author(s):

Nuschin Morakkabati-Spitz ◽

Jürgen Gieseke ◽

Winfried A. Willinek ◽

Patrick J. Bastian ◽

Bettina Schmitz ◽

...

Keyword(s):

Urinary Incontinence ◽

Mr Imaging ◽

Pelvic Floor ◽

Clinical Signs ◽

Preliminary Results

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Hepatozoon canis and Leishmania spp. coinfection in dogs diagnosed with visceral leishmaniasis

Revista Brasileira de Parasitologia Veterinária ◽

10.1590/s1984-29612016065 ◽

2016 ◽

Vol 25 (4) ◽

pp. 450-458 ◽

Cited By ~ 6

Author(s):

Fernanda Nazaré Morgado ◽

Amanda dos Santos Cavalcanti ◽

Luisa Helena de Miranda ◽

Lúcia Helena O’Dwyer ◽

Maria Regina Lucas da Silva ◽

...

Keyword(s):

Visceral Leishmaniasis ◽

Dna Isolation ◽

Parasite Burden ◽

Leishmania Species ◽

18S Rrna Gene ◽

Hepatozoon Canis ◽

Histopathological Analysis ◽

Rrna Gene ◽

Pcr Rflp ◽

Leishmania Spp

Abstract This study describes the occurrence of dogs naturally co-infected with Hepatozoon canis and two Leishmania species: L. infantum or L. braziliensis. Four dogs serologically diagnosed with Visceral Leishmaniasis were euthanized. Liver and spleen samples were collected for histopathological analysis and DNA isolation. H. canis meronts were observed in tissues from all four dogs. H. canis infection was confirmed by PCR followed by sequencing of a fragment of 18S rRNA gene. Leishmania detection and typing was confirmed by ITS1' PCR-RFLP and parasite burden was calculated using ssrRNA quantitative qPCR. A DPP - Dual Path platform test was performed. One out (Dog #2) of four animals was asymptomatic. Dogs #1 and #4 were infected by L. infantum and were DPP test positive. Dogs #2 and #3 were infected by L. braziliensis and were DPP test negative. Furthermore, visceral dissemination was observed in Dogs #2 and #3, since L. braziliensis was detected in liver and spleen samples. The visceral dissemination of L. braziliensis associated with systemic signs suggested that this co-infection could influence the parasite burden and disease progression.

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